Microbiological,compositional, biochemical and textural characterisation of Caciocavallo Pugliese cheese during ripening

A microbiological, compositional, biochemical and textural characterisation of the pasta filata Caciocavallo Pugliese cheese during ripening is reported. Fully ripened cheese contained a total of ca. log 8.0 cfu g⁻¹ mesophilic bacteria and ca. log 6.0 cfu g⁻¹ presumptive staphylococci, while the number of thermophilic and mesophilic rod and coccus lactic acid bacteria varied during ripening. A two-step RAPD-PCR protocol was used to differentiate biotypes. The natural whey starter was composed mainly of Lactobacillus delbrueckii, Lb. fermentum, Lb. gasseri, Lb. helveticus and Streptococcus thermophilus strains. After day 1 of ripening, Lb. delbrueckii became dominant and some strains of Enterococcus durans and E. faecalis appeared. Non-starter lactic acid bacteria, such as Lb. parabuchneri and Lb. paracasei subsp. paracasei formed a large part of the lactic microflora at 42 and 60 d of ripening. The level of pH 4.6-soluble nitrogen increased from the outer to the inner of the cheese and also increased in each section as ripening progressed, attaining values of 18–15%. Urea-PAGE electrophoresis showed that degradation of α_s1-casein was more rapid than that of β-casein throughout ripening and the rates at which both caseins were degraded greatly increased from the outside to the inside of the cheese. Based on the primary proteolysis products, both chymosin and plasmin appeared to be active. RP-HPLC profiles of the 70% ethanol-soluble, pH 4.6-soluble nitrogen, showed a large number of peaks, indicating a heterogeneous mixture of proteolytic products. There were both age- and section-related changes in the area of the different peptide peaks. Butyric (C4:0), caproic (C6:0), palmitic (C16:0) and oleic (C18:1) acids were the free fatty acids found at the highest concentrations. The level of short chain fatty acids (e.g., butyric and caproic) decreased from the middle and inner to outer sections of the cheese. Peptidase activity in the curd was pronounced, increased during ripening and varied with the cheese section. The greatest increase of the peptidase activity coincided with a change in the lactic microflora and with the prevalence of non-starter lactic acid bacteria. Microbial esterases were supposed to be active together with rennet paste. Little change in the firmness and fractures stress during maturation were found by textural analyses of the raw cheese. The flowability was similar to that of typical low-moisture Mozzarella cheese, while stretchability was lower. The heat-induced changes in phase angle of Caciocavallo Pugliese cheese indicated a phase transition from largely elastic rheological characteristics in unheated cheese to a more viscous and fluid character in melted cheese.     

Effects of Commercial Food Grade Enzymes on Free Fatty Acid Profiles in Granular Cheddar Cheese

Commercial food grade enzymes (Neutrase, calf lipase and NaturAge) were incorporated into cheese at various concentrations and ripened at 7° and 13°C. Gas chromatographic analysis indicated that free fatty acid (FFA) production increased significantly (p<0.05) with lipase and high level NaturAge. Neutrase had little effect on FFA production but showed synergistic effects with lipase. The short-chain FFA profiles were similar among control, Neutrase- and NaturAge-treated cheeses. Significant correlations (p<0.05) between C₈ and C₁₄, C₁₆ or C_18:1 and total free fatty acids were observed in all tested samples. Total concentrations of C₄ and C₆ may be a good indicator of flavor development during cheese ripening. Cheese with a medium level of NaturAge, ripened at 13°C for 4 wk, showed profiles of FFA similar to those of the control cheese ripened at 7°C for 12 wk.     

Free fatty acids in model cheeses made with a Micrococcus sp. INIA 528 milk culture or with a high enzymatic activity curd of this strain

The effect of milk inoculation with cultures of Micrococcus sp. INIA 528, and addition of high enzymatic activity curd prepared with this strain to standard curd made with a lactic starter culture, on the release of free fatty acids during ripening of model cheeses was investigated. Milk inoculation with the Micrococcus culture had no significant effect on free fatty acid levels. Addition of high enzymatic activity curd to standard curd resulted in levels of total free fatty acids (C4–C18) 5.4-, 3.9- and 5.0-fold more than those in control cheese on days 1, 7 and 14, respectively, when added at a level of 20%, and 2.9-, 2.9- and 3.1-fold when added at a level of 10%. The highest increases in individual free fatty acids in cheeses made with high enzymatic activity curd corresponded to hexanoic, butanoic and octanoic acids.     

The technological characteristics of Debaryomyces hansenii and Yarrowia lipolytica and their potential as starter cultures for production of Danablu

Six strains of Debaryomyces hansenii var. hansenii and Yarrowia lipolytica, respectively, originating from blue mould cheeses were examined for their potential use as starter cultures for the production of Danablu in laboratory studies. D. hansenii showed strong growth and assimilation of lactose, galactose, lactate and five out of six strains assimilated citric acid under the environmental conditions prevailing in Danablu during maturation at 10°C. Y. lipolytica was more sensitive to NaCl and did not assimilate lactose and galactose. Both yeasts hydrolysed tributyrin with the highest activity observed for Y. lipolytica. D. hansenii showed little if any release of free fatty acids from butterfat at 10°C. Y. lipolytica was strongly lipolytic. The strains of D. hansenii were not able to hydrolyse casein at 10°C whereas 4 of the 6 strains of Y. lipolytica degraded all components of the casein. Strain-specific interactions, in cheese agar resulting in inhibition of mycelial growth and sporulation of P. roqueforti was observed, especially for Y. lipolytica.     

High-Pressure Processing for the Control of Lipolysis,Volatile Compounds and Off-odours in Raw Milk Cheese

Build-up of flavour compounds throughout ripening of raw milk cheeses may result in strong over-ripening notes during refrigerated storage. In order to control the formation of free fatty acids (FFAs) and volatile compounds, and the appearance of off-odours, raw milk cheeses were high-pressure-processed (HPP) 21 or 35 days after manufacture at 400 or 600 MPa. Ripening proceeded at 8 °C until day 60 and, afterwards, cheeses were held at 4 °C until day 240. The effect of HPP on the formation of FFAs and volatile compounds was dependent on pressure level and cheese age at the time of treatment. On day 60, acetic and propionic acids, branched-chain FFAs and short-chain FFAs showed the lowest (p?<?0.05) concentrations in cheeses treated at 400 or 600 MPa on day 21, while medium- and long-chain FFAs were at similar levels in all cheeses. HPP influenced significantly (p?<?0.05) 84 out of the 94 volatile compounds found in cheese. On day 60, the lowest (p?<?0.05) concentrations of acids, alcohols and esters were recorded for cheeses treated at 400 or 600 MPa on day 21, and the lowest (p?<?0.05) concentrations of ketones for cheeses treated at 400 MPa on days 21 or 35. On day 240, all HPP cheeses showed lower (p?<?0.05) concentrations of aldehydes, esters and, particularly, sulphur compounds than control cheese, which exhibited putrid and rancid off-odours from day 120 onwards. Principal component analysis combining FFAs and volatile compounds discriminated 240-day control cheese from 120-day control cheese and both from the rest of cheeses.     

Evolution of lipolysis during the ripening of traditional Feta cheese

Lipolysis was studied during ripening of traditional Feta cheese produced in two small dairies, A and B. The cheeses were made from a thermized mixture of ewes’/goats’ milk by using yoghurt as starter and artisanal rennet from lambs’ and kids’ abomasa (cheese A) or mixed artisanal rennet with calf rennet (cheese B).The acid degree value and the free fatty acids (FFA) contents in both cheeses increased sharply up to 18 d (pre-ripening period at 15 °C) and continued to increase throughout ripening. In both mature cheeses, acetic acid was found at high levels (13–18% of the total FFAs). However, except for this, all FFA contents differed significantly (P < 0.05) between the two cheeses throughout ripening. The levels of individual and total C2:0–C8:0, C10:0–C14:0 and C16:0–C18:2 fatty acids were significantly higher (P < 0.05) in cheese A than in cheese B. Presumably the difference, especially in the C2:0–C8:0 content, was due mainly to the type of the rennet used. Butyric acid was the dominant FFA in cheese A (20% of the total FFAs at 120 d), while the most abundant FFAs in cheese B were capric (18%) and lauric acid (18%). In general, the lipolysis degree of the two cheeses was higher than those reported for the industrially-made Feta cheese.In organoleptic evaluation, cheese A had a piquant taste that was attributed to its high content of butyric acid and showed a significantly (P < 0.05) higher total score than cheese B.     

Evolution of Free Amino Acids during the Ripening of Cheddar Cheese Containing Added Lactobacilli Strains

Cheddar cheese was produced with different lactobacilli strains added to accelerate ripening. The concentration of proteolytic products was determined as free amino acids in the water-soluble fraction at two, four, seven and nine months of aging and at two different maturation temperatures (6°C, 15°C). All amino acids increased during ripening and were higher in the Lactobacillus- added cheeses than in the control cheese, and higher in cheeses ripened at 15°C than at 6°C. Glutamic acid, leucine, phenylalanine, valine and lysine were generally in higher proportion in all cheeses. The cheeses with added L. casei-casei L2A were classified as having a “strong Cheddar cheese” flavor after only seven months of ripening at 6°C.     

Effects of heating milk and accelerating ripening of low fat Ras cheese on biogenic amines and free amino acids development

Two experiments were conducted to study the effects of heating milk and, consequently, incorporation of whey proteins into cheese curd, fat content, accelerating cheese ripening by attenuated lactobacilli, species of bacteria and method of attenuation on formation of biogenic amines and liberation of free amino acids. Total biogenic amines and total free amino acids increased as ripening period progressed in all cheese treatments. Total biogenic amines and total free amino acids decreased as the fat content was decreased. Heating of 3% fat milk up to 70°C caused a significant (p<0.05) increase in total biogenic amines and total free amino acid concentrations, while raising the temperature of heat treatment up to 75 and 80°C decreased them. However, heating of 2% fat milk up to 75°C caused a definite (p<0.05) increase in total biogenic arnines and total free amino acids; conversely raising heat treatment temperature to 80°C decreased them. These results indicate that there is a positive correlation between total biogenic amines and total free amino acids; moisture and salt contents affected the formation of biogenic amines, while incorporation of whey proteins had no significant effect on biogenic amines development. In a second experiment, addition of attenuated lactobacilli as adjunct bacteria caused a pronounced (p<0.05) increase in free amino acids and biogenic amines. Addition of either freeze- or heat-shocked Lactobacillus helviticus was more effective in promoting the build-up of biogenic amines and free amino acids in the resultant cheeses than in cheeses made with addition of freeze- or heat-shocked Lactobacillus casei, respectively. There was a positive correlation between free amino acids and biogenic amine contents. The type of bacteria and prolongation of ripening period significantly affected the development of biogenic amines. Tyramine was the highest biogenic amine followed by histamine, while spermidine was the lowest.     

Evaluation of metabolic activities of enterococci isolated from Pecorino Abruzzese cheese

Enterococci, which represent a considerable part of the natural microbiota of Pecorino Abruzzese, were investigated for biochemical activities considered to have a potential role in cheese ripening. Acidifying activity was weak, while interesting differences were identified for proteolytic and peptidolytic capability, particularly on H-Lys-β-NA. The presence of C-4 esterase and C-8 esterase lipase, as well as enzymes such as phosphohydrolases and acid phosphatase was confirmed by means of API ZYM. SPME-GC analyses of volatile compounds revealed the production of ethanol, diacetyl and, of particular interest, acetoin after 15 days at 10 °C, with important differences among the species. After 15 days at 10 °C, Enterococcus faecalis isolates were generally the most active, giving the highest results for most of the biochemical activities investigated, and also some Enterococcus durans showed interesting metabolic features that might be important to the development of the peculiar sensory properties of this cheese.     

Determination of lipase activity in cheese using trivalerin as substrate

Lipolysis during cheese ripening is usually assessed by the accumulation of free fatty acids (FFA). An assay to determine total lipolytic activity present in a cheese during ripening was established. Finely grated cheese (1 g) was directly incubated with trivalerin (204 mg) as a substrate at 35 °C for 4 h. Free valeric acid was extracted and quantified by gas chromatography. The assay was linear with time up to 24 h and up to 0.11 Lipase Units g⁻¹ cheese. The total amount of lipolytic activity determined with this assay was consistent with the total amount of FFA present in 5 types of cheese. In Idiazabal cheese samples manufactured with or without lipase added, the total lipolytic activity determined during ripening remained constant up to 6 months of ripening. In addition to trivalerin, other possible substrates investigated were triolein, triundecanoin and the endogenous butyric acid-containing triacylglycerols present in the cheese sample. Activities measured with these substrates were considerably lower than values obtained with trivalerin due to the high levels of oleic acid present in cheese, or to difficulties in mixing triundecanoin (solid at 35 °C) with the grated cheese sample. Endogenous triacylglycerols gave increasingly lower activity values as ripening time progressed due to substrate depletion.     

The isolation and identification of yeasts obtained during the manufacture and ripening of Cheddar cheese

Sources of yeast, which may contaminate the curd during the manufacture of Cheddar cheese, were examined in a single cheese factory. A total of 77 yeast species present in the factory environment, manufacturing and ripening of Cheddar cheese were identified according to cellular long-chain fatty acid analysis and verified with conventional identification techniques. Product line samples were taken at critical control points in the manufacturing process and analysed after incubation at 25°C for 96 h. The progression of yeast species during cheese-making and ripening was monitored after renneting and at subsequent 48-h intervals. Dominant species wereDebaryomyces hanseniiandCryptococcus albidus, whileYarrowia lipolytica, Rhodotorula minuta, Torulaspora delbrueckii, Rhodotorula glutinisandKluyveromyces marxianuswere present at low numbers. The results obtained showed that yeasts were present in all cheese samples examined, at quantities ranging from 9×10²to 1·4×10⁷cfu g⁻¹.     

Effect of ripening temperature on the growth and significance of non-starter lactic acid bacteria in Cheddar cheese made from raw or pasteurised milk

Two cheese-making trials were conducted, each involving four cheeses, two made from raw milk (R1, R8) and two from pasteurised milk (P1, P8), and ripened at 1°C (R1, P1) or 8°C (R8, P8). The 1-day-old R1 and R8 cheese in trials 1 and 2 contained ∼10⁴ non-starter lactic acid bacteria (NSLAB) g⁻¹. In trial 1, no NSLAB were detected in 1-day-old P1 and P8 cheeses while those in trial 2 contained 10² cfu g⁻¹. In both trials, the maximum differences between the number of NSLAB in the cheeses ripened at 1 or 8°C were observed at 4 months, when the number of NSLAB in cheeses ripened at 8°C were 3 log cycles higher than in those ripened at 1°C. At the end of ripening (6-months), the number of NSLAB in P8 and R8 were ∼2 log cycles higher than in P1 and R1 cheeses, respectively. Primary proteolysis in the cheeses was markedly affected by ripening temperature, but not by pasteurisation of the cheese milk. Urea-polyacyrlamide gel electrophoretograms and reverse-phase (RP)-HPLC of the water-soluble fraction showed differences between cheeses made from raw or pasteurised milk and between cheeses ripened at 1 or 8°C. The concentration of amino acids and fatty acids were in the order R8>P8>R1>P1. Commercial graders awarded highest flavour scores to the R1 cheeses during gradings at 4, 5 and 6 months. A sensory panel found that most flavour and aroma attributes and maturity were in the order of R8>P8>R1=P1. The results of this study suggest that NSLAB play an important role in the development of flavour in Cheddar cheese by contributing to the production of amino acids and fatty acids.     

β-Galactosidase in the acceleration of Ras cheese ripening

An attempt has been carried out to accelerate Ras cheese ripening by pre-treatment of cheese milk with β-galactosidase. Milk was treated with a β-galactosidase enzyme preparation, namely lactozym (1 ml/kg milk), at 33°C for 1 h or at 4°C for 18 h and used for Ras cheese making. Flavour intensity, formation of soluble nitrogen compounds, free amino acids and liberation of free fatty acids were enhanced in cheese made from β-galactosidase treated milk. In addition, the ripening period was reduced to 2 months compared with 4 months required for control cheese. Treatment of cheese milk with β-galactosidase at 4°C or 33°C showed a similar effect on the properties of cheese.     

Esterase activities of indigenous lactic acid bacteria from Argentinean goats’ milk and cheeses

Esterase activities of indigenous lactic acid bacteria isolated from goats’ milk and cheese were investigated. All of the strains exhibited esterase activity on α-naphthyl derivative of fatty acids of 2–6 carbon atoms. Lactobacillus fermentum ETC1 and L. bulgaricus ETC2 showed the highest specific activity on α-naphthyl acetate; L. rhamnosus ETC14 presented the highest specific activity on α-naphthyl butyrate and caproate. All enterococci strains presented the highest specific activities on α-naphthyl propionate, butyrate and caproate, and the lowest specific activities on α-naphthyl acetate. Pedicoccus pentocaseus ETC5 only had esterase activity on α-naphthyl acetate. The electrophoretic zymogram showed for each strain an individual enzyme profile on α-naphthyl acetate and revealed the presence of more than one esterase. L. plantarum ETC11 and Enterococcus faecium ETC9 showed four and five bands of esterase activity, respectively. The strains evaluated in this work showed different esterase activities, which were species and strain specific.     

Freezing low moisture Mozzarella cheese: changes in organic acid content

Vacuum-packaged low moisture Mozzarella cheese samples were ripened both by the traditional method at 4°C and after subjecting the samples to a freeze (−20°C)–thaw (4°C) cycle before ageing. Nine organic acids (formic, pyruvic, orotic, uric, lactic, acetic, citric, propionic and butyric) were analyzed each week by high performance liquid chromatography. There was no significant effect of the freeze–thaw cycle on the variations in organic acids content. They were only affected by ripening time. Each organic acid presented a characteristic pattern of change during ripening. Discriminant analysis classified cheeses according to their age.     

Biochemical patterns in ovine cheese: Influence of probiotic strains

This study was undertaken to evaluate the effect of lamb rennet paste containing probiotic strains on proteolysis, lipolysis, and glycolysis of ovine cheese manufactured with starter cultures. Cheeses included control cheese made with rennet paste, cheese made with rennet paste containing Lactobacillus acidophilus culture (LA-5), and cheese made with rennet paste containing a mix of Bifidobacterium lactis (BB-12) and Bifidobacterium longum (BB-46). Cheeses were sampled at 1, 7, 15, and 30 d of ripening. Starter cultures coupled with probiotics strains contained in rennet paste affected the acidification and coagulation phases leading to the lowest pH in curd and cheese containing probiotics during ripening. As consequence, maturing cheese profiles were different among cheese treatments. Cheeses produced using rennet paste containing probiotics displayed higher percentages of α_S1-I-casein fraction than traditional cheese up to 15 d of ripening. This result could be an outcome of the greater hydrolysis of α-casein fraction, attributed to higher activity of the residual chymosin. Further evidence for this trend is available in chromatograms of water-soluble nitrogen fractions, which indicated a more complex profile in cheeses made using lamb paste containing probiotics versus traditional cheese. Differences can be observed for the peaks eluted in the highly hydrophobic zone being higher in cheeses containing probiotics. The proteolytic activity of probiotic bacteria led to increased accumulation of free amino acids. Their concentrations in cheese made with rennet paste containing Lb. acidophilus culture and cheese made with rennet paste containing a mix of B. lactis and B. longum were approximately 2.5 and 3.0 times higher, respectively, than in traditional cheese. Principal component analysis showed a more intense lipolysis in terms of both free fatty acids and conjugated linoleic acid content in probiotic cheeses; in particular, the lipolytic pattern of cheeses containing Lb. acidophilus is distinguished from the other cheeses on the basis of highest content of health-promoting molecules. The metabolic activity of the cheese microflora was also monitored by measuring acetic, lactic, and citric acids during cheese ripening. Cheese acceptability was expressed for color, smell, taste, and texture perceived during cheese consumption. Use of probiotics in trial cheeses did not adversely affect preference or acceptability; in fact, panelists scored probiotic cheeses higher in preference over traditional cheese, albeit not significantly.     

Integrated Roles of Lactate,Ammonia, and Calcium in Texture Development of Mold Surface-Ripened Cheese

Pure cultures of Penicillium caseicolum and Geotrichum candidum each rapidly metabolized lactic acid from an acidified milk-agar medium. Penicillium caseicolum rapidly produced ammonia on milk agar (within 1 wk) but G. candidum elaborated ammonia only in older cultures (>3 wk). Brevibacterium linens showed little metabolic activity on milk agar systems because of inhibition by low pH or lack of proteases or essential nutrients. Analysis of samples (center, corner, surface locations) from wheels of a lot of ripening Brie cheese (227 g each) held at 17°C over 57 d also showed metabolic depletion of lactic acid and the production of ammonia by the surface microflora. During ripening, calcium concentrations became elevated in surface locations and diminished in interior locations of Brie wheels. An integrated model is proposed that incorporates a pH mediation of systems for calcium migration and insolubilization, proteolytic enzyme activity, and casein solubilization whose combined activities control texture development in mold surface-ripened cheeses.     

Proteolytic and lipolytic changes during the ripening of a Spanish craft goat cheese (Armada variety)

The proteolytic and lipolytic changes during the ripening process were investigated in four batches of Armada goat's milk cheese (an artisanal variety produced in the North of Spain), by determining the classical nitrogen fractions, caseins and their degradation products, free amino acids, as well as the acidity of the fat, thiobarbituric acid (TBA) number and free fatty acids. Values obtained for the nitrogen fractions and for caseins and their degradation products show that this cheese undergoes very little protein degradation. A low free amino acids content was observed throughout the ripening process with a predominance of Pro followed by Leu+Ile, Glu acid, Phe, His+Lys and Val. The lipid degrada-tion was very intense from the second month of ripening, only comparable to that reported for cheeses ripened by moulds. The average free fatty acids content increased 20-fold during ripening, reaching final values of 44·5 g kg⁻¹. All the free fatty acids increased considerably during ripening, resulting in a predominance of saturated and unsaturated long-chain acids, followed by medium-chain acids, C₁₀ principally. Short-chain fatty acid content by the end of ripening was higher than that presented in other cheese varieties with a similar high degree of lipolysis. © 1997 SCI.     

Effect of starter cultures on the physico‐chemical and biochemical features in ewe cheese made with extracts from flowers of Cynara cardunculus L

The effect of starter cultures on the physico‐chemical and biochemical features of Los Pedroches cheese made by using Cynara Cardunculus L extracts as vegetable coagulant was assessed. Specimens of cheese containing no starter (vats A and C) and others to which a lactic acid starter was added (vats B and D) were stored either under typical farmhouse ripening conditions (vats A and B) or in a controlled chamber at 14°C and 80% relative humidity (vats C and D) for 90 days. The addition of starter cultures and the ripening conditions where found to have no appreciable effect on the moisture, fat, protein, ash, NaCl, Ca and P contents of the cheese, nor on its water activity; however, it resulted in increased lactic acid contents and lower pH values relative to control specimens throughout ripening. The protein breakdown of the cheeses was assessed in terms of soluble nitrogen, non‐protein nitrogen, aminoacid nitrogen, ammonia nitrogen, soluble tyrosine and tryptophan, and urea‐PAGE. Proteolysis was generally more marked and rapid in cheese containing the lactic starter, the NPN, NH₃ ‐N and soluble Tyr and Trp contents of which were significantly higher than those of the cheese obtained with no starter. The PAGE technique revealed no appreciable differences between the four cheese batches studied; however, it showed the cheese to be more resistant to β‐CN hydrolysis than to α_s‐CN hydrolysis. In general, the flavour and aroma scores were higher in batches obtained with starter cultures and ripened under controlled conditions. © 1999 Society of Chemical Industry     

Behavior of Shiga-toxin-producing Escherichia coli in ewe milk stored at different temperatures and during the manufacture and ripening of a raw milk sheep cheese (Zamorano style)

This study was conducted to assess the survival of 2 wild Shiga toxin-producing Escherichia coli strains (one serotype O157:H7 and one non-O157:H7) in ewe milk stored at different conditions and to examine the fate of the O157 strain during the manufacture and ripening of a Spanish sheep hard variety of raw milk cheese (Zamorano). The strains were selected among a population of 50 isolates, which we obtained from ewe milk, because of their high resistance to 0.3% lactic acid. Both strains were inoculated (approximately 2 log₁₀ cfu/mL) in raw and heat-treated (low-temperature holding, LTH; 63°C/30 min) ewe milk and stored for 5 d at 6, 8, and 10°C and also according to a simulation approach for assessing the effects of failures in the cold chain. The minimum growth temperature for the O157:H7 strain in LTH and raw ewe milk was 8°C. For the non-O157:H7 strain, the lowest temperature showing bacterial growth in LTH ewe milk was 6°C, but it did not grow at any of the tested conditions in raw milk. It appears that the O157 strain was more susceptible to cold stress but was likely a better competitor than the non-O157 strain against the milk autochthonous microbiota. For manufacture of Zamorano cheese, raw milk was inoculated with approximately 3 log₁₀ cfu/mL, and after 2 mo of ripening at 10 to 12°C, the cheeses showed the expected general characteristics for this variety. The O157:H7 strain increased 0.9 log₁₀ cfu/g after whey drainage and during ripening and storage decreased by 2.9 log₁₀ cfu/g. Nevertheless, its detectable level (estimated at 6.2 cfu/g) after 2 mo of ripening suggests that Zamorano cheese manufactured from raw ewe milk contaminated with E. coli O157:H7 could represent a public health concern.